Type of Novel Lipid Compound and Use Thereof

This application is a U.S. National Stage Application of PCT Application No. PCT/CN2023/095573, filed on May 22, 2023, which claims priority to Chinese Patent Application No. 202210679273.8, filed on Jun. 16, 2022, the entire contents of which are hereby incorporated by reference in their entirety for all purpose.

The present disclosure belongs to the field of biomedicine and biotechnology, and relates to a novel lipid compound and a system for constructing a lipid nanodelivery carrier by using the lipid compound to deliver active ingredients.

When mRNA drugs are used in clinical treatment, it is necessary to overcome many obstacles in the delivery of exogenous mRNA, and therefore, safe and effective carriers are needed to deliver mRNA to target tissues, organs and cells in the body to play a corresponding role. Lipid nanoparticles (LNPs) are currently the most advanced mRNA delivery system, which are safe and efficient and represents the mainstream of mRNA carrier development in the future.

Lipid nanoparticles (LNPs) are a mature delivery platform for nucleic acids, generally including the nucleic acid that are desired to be delivered, cationic/ionizable/lipoid and some auxiliary lipids, wherein the auxiliary lipids are usually phospholipids, cholesterol and PEGylation lipids. LNPs are currently the most advanced nanodrug delivery system for nucleic acid drugs. How to prepare stable, safe and delivery-efficient LNPs, achieve rapid conversion of gene drugs and achieve targeted delivery to different tissues are key issues in this field, and the solution to these issues depends on a lipid molecule library with diverse structures and functions.

Ionizable lipids (ILs), also called pH-dependent lipids, are almost uncharged and neutral at physiological pH. Under acidic conditions, ILs are positively charged, which facilitates assembly with negatively charged mRNA by electrostatic interactions. ILs remain neutral in the neutral environment of the body fluids. ILs are protonated as the pH decreases below the pKa of the ILs during cellular internalization, and due to the proton sponge effect, LNPs osmotically swell and rupture, releasing mRNA. The chemical structure of ILs plays a decisive role in factors such as the stability, biosafety, and delivery efficiency of LNPs.

The ionizable lipid structure generally comprises three moieties: a hydrophilic head, a hydrophobic tail, and a linker moiety connecting the head and the tail. Based on current research progress and clinical conditions, degradable and multibranched tails are favorable structural properties for the future development of ionizable lipids. For example, the structure of the ionizable lipid SM-102 used by Moderna in the COVID-19 vaccine includes a tertiary amine head, three branches, and a tail containing an ester bond. As the most critical component in LNPs, screening for safer and more efficient ionizable lipids has always been the focus of improving the performance of LNPs.

The present disclosure provides a type of novel lipid compound with a simple preparation method, low toxicity and high biocompatibility, which enriches the types of lipid compounds and provides more choices for the delivery of nucleic acid drugs. The lipid compound of the present disclosure, when prepared into LNPs with other lipids, is capable of effectively delivering mRNA or drug molecules into cells to exert biological functions.

The present disclosure provides a compound of formula (I), or a salt or an isomer thereof, wherein the formula (I) has a structure as shown below:

In some embodiments, Rin the compound of formula (I) is selected from —CHCH, —CHCHCH, —CH(CH), —CHCH(CH), —(CH)CH, —C(CH), —CH(CH)CHCH, —CHCHOH, —CH(OH)CH, —CHCHCHOH, —CHCH(CH)OH, —CH(CH)CHOH, —C(OH)(CH), —CH(OH)CHCH, —CHN(CHCH), —CHN(CH), —CHNHCH, —CHNHCHCH, —CHN(CH)CHCH, —CH(OCHCH),

wherein Ris selected from Calkyl, Calkoxy or halogen, and p is selected from a natural number of 0-2.

In some embodiments, Rin the compound of formula (I) is selected from —CHCHCH, —CHCH(CH), —CHCHOH, —CHCHCHOH, —CHCH(CH)OH, —CH(OH)CHCH, —CHN(CHCH), —CHNHCH, —CHN(CH), —CH(OCHCH),

In some embodiments, Rin the compound of formula (I) is selected from —CHCHOH, —CHCHCHOH, —CHCH(CH)OH, —CH(OH)CHCH, —CHN(CHCH), —CHN(CH),

In some embodiments, Rin the compound of formula (I) is selected from —CHN(CHCH), —CHN(CH),

In some embodiments, Rin the compound of formula (I) is selected from Calkyl or Calkenyl.

In some embodiments, Rin the compound of formula (I) is selected from Calkyl or Calkenyl.

In some embodiments, Rin the compound of formula (I) is selected from —(CH)CH, (CH)CH, —(CH)CH, —(CH)CH, —(CH)CH, —(CH)CH, —CH((CH)CH), —CH((CH)CH), —CH((CH)CH), —CH((CH)CH), —CH((CH)CH), (CH)CH═CH(CH)CH, —(CH)CH═CH(CH)CH, —(CH)CH═CH(CH)CH, (CH)CH═CH(CH)CH, —(CH)CH═CH(CH)CH, —(CH)CH═CH(CH)CH, (CH)CH═CH(CH)CH, —(CH)CH═CH(CH)CH, —(CH)CH═CH(CH)CH, (CH)CH═CH(CH)CH, —(CH)CH═CH(CH)CH, —(CH)CH═CH(CH)CH, (CH)CH═CH(CH)CH, —(CH)CH═CH(CH)CH, —(CH)CH═CH(CH)CH, (CH)CH═CH(CH)CH, —(CH)CH═CH(CH)CH, —(CH)CH═CH(CH)CH, (CH)CH═CH(CH)CHor —(CH)CH═CH(CH)CH.

In some embodiments, Rin the compound of formula (I) is selected from —(CH)CH, (CH)CH, —(CH)CH, —(CH)CH, —(CH)CH, —(CH)CH, —CH((CH)CH), —CH((CH)CH), —CH((CH)CH), —CH((CH)CH), —CH((CH)CH).

In some embodiments, Rin the compound of formula (I) is selected from —(CH)CH, (CH)CH, —(CH)CH, —(CH)CH, —(CH)CH, —(CH)CH, —CH((CH)CH), —CH((CH)CH), —CH((CH)CH), —CH((CH)CH), (CH)CH═CH(CH)CH, —(CH)CH═CH(CH)CH, —(CH)CH═CH(CH)CH, (CH)CH═CH(CH)CH, —(CH)CH═CH(CH)CH, —(CH)CH═CH(CH)CH, (CH)CH═CH(CH)CH, —(CH)CH═CH(CH)CH, —(CH)CH═CH(CH)CH, (CH)CH═CH(CH)CH, —(CH)CH═CH(CH)CH, —(CH)CH═CH(CH)CH, (CH)CH═CH(CH)CH, —(CH)CH═CH(CH)CH, —(CH)CH═CH(CH)CH, (CH)CH═CH(CH)CH, —(CH)CH═CH(CH)CH, —(CH)CH═CH(CH)CH, —CH((CH)CH), (CH)CH═CH(CH)CHor —(CH)CH═CH(CH)CH.

In some embodiments, Rin the compound of formula (I) is selected from —(CH)CH, (CH)CH, —(CH)CH, —(CH)CH, —(CH)CH, —(CH)CH, —CH((CH)CH), —CH((CH)CH), —CH((CH)CH), —CH((CH)CH), —CH((CH)CH).

In some embodiments, Rand Rin the compound of formula (I) are independently selected from —(CH)CH, —(CH)CH, —(CH)CH, —(CH)CH, —(CH)CH, —(CH)CH, —CH((CH)CH), —CH((CH)CH), —CH((CH)CH), —CH((CH)CH), —CH((CH)CH),

In some embodiments, n and m in the compound of formula (I) are each independently selected from an integer of 3-9. In some embodiments, n and m in the compound of formula (I) are each independently selected from an integer of 4-8. In some embodiments, n and m in the compound of formula (I) are each independently selected from an integer of 5-7. In some embodiments, n and m in the compound of formula (I) are each independently selected from an integer of 5, 6 or 7.

In some embodiments, in the compound of formula (I), n is 5, m is 7, Ris —(CH)CH, and Ris —CH((CH)CH).

In some embodiments, in the compound of formula (I), both n and m are 7, and both Rand Rare —CH((CH)CH).

In some embodiments, in the compound of formula (I), n is 5, m is 7, Ris (CH)CH═CH(CH)CH, and Ris —CH((CH)CH).

In some embodiments, the compound of formula (I), or the salt or the isomer thereof is selected from the following compounds LipidA-1 to LipidA-14, LipidB22-1 to LipidB22-15, LipidB23-1 to LipidB23-15, or salts or isomers thereof.

The present disclosure further provides a delivery carrier, comprising the compound of the present disclosure and an accessory molecule. In some embodiments, the accessory molecule comprises: a phospholipid, a structural lipid and a PEGylation lipid.

In some embodiments, in the delivery carrier of the present disclosure, the molar ratio of the compound of the present disclosure to the accessory molecule is 1:1.

In some embodiments, in the delivery carrier of the present disclosure, the content of the compound of the present disclosure is 20%-80%, the content of the PEGylation lipid compound is 1%-10%, the content of the structural lipid is 10%-50%, and the content of the phospholipid is 5%-30%, by mole percent. Optionally, the content of the compound of formula (I) is selected from 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75% or 80% by mole percent. Optionally, the content of the compound of formula (I) is selected from 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54% or 55% by mole percent. Optionally, the content of the PEGylation lipid compound is selected from 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5% or 10% by mole percent. Optionally, the content of the PEGylation lipid compound is selected from 1%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9% or 2% by mole percent. Optionally, the content of the structural lipid is 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45% or 50% by mole percent. Optionally, the content of the structural lipid is 35%, 35.5%, 36%, 36.5%, 37%, 37.5%, 38%, 38.5%, 39%, 39.5% or 40% by mole percent. Optionally, the content of the phospholipid is selected from 5%, 10%, 15%, 20%, 25% or 30% by mole percent. Optionally, the content of the phospholipid is 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14% or 15% by mole percent.

In some embodiments, the phospholipid is selected from at least one of 1,2-dilinoleoyl-sn-glycero-3-phosphocholine (DLPC), 1,2-dimyristoyl-sn-glycero-phosphocholine (DMPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-diundecanoyl-sn-glycero-phosphocholine (DUPC), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), 1,2-di-O-octadecenyl-sn-glycero-3-phosphocholine (18:0Diether PC), 1-oleoyl-2-cholesterylhemisuccinoyl-sn-glycero-3-phosphocholine (OChemsPC), 1-hexadecyl-sn-glycero-3-phosphocholine (C16 Lyso PC), 1,2-dilinolenoyl-sn-glycero-3-phosphocholine, 1,2-diarachidonoyl-sn-glycero-3-phosphocholine, 1,2-didocosahexaenoyl-sn-glycero-3-phosphocholine (DOPE), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (ME 16.0PE), 1,2-diphytanoyl-sn-glycero-3-phosphoethanolamine 1,2-distearoyl-sn-glycero-3-phosphoethanolamine, 1,2-dilinoleoyl-sn-glycero-3-phosphoethanolamine, 1,2-dilinolenoyl-sn-glycero-3-phosphoethanolamine, 1,2-diarachidonoyl-sn-glycero-3-phosphoethanolamine, 1,2-didocosahexaenoyl-sn-glycero-3-phosphoethanolamine, 1,2-dioleoyl-sn-glycero-3-phospho-rac-(1-glycerol) sodium salt (DOPG), dipalmitoyl phosphatidylglycerol (DPPG), palmitoyl oleoyl phosphatidylethanolamine (POPE), distearoyl-phosphatidylethanolamine (DSPE), dipalmitoyl phosphatidylethanolamine (DPPE), dimyristoyl phosphatidylethanolamine (DMPE), 1-stearoyl-2-oleoyl-stearoylethanolamine (SOPE), 1-stearoyl-2-oleoyl-phosphatidylcholine (SOPC), sphingomyelin, phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, phosphatidic acid, palmitoyloleoylphosphatidylcholine, lysophosphatidylcholine or lysophosphatidylethanolamine (LPE).

In some embodiments, the structural lipid is selected from at least one of cholesterol, coprosterol, sitosterol, ergosterol, campesterol, stigmasterol, brassicasterol, tomatine, ursolic acid or α-tocopherol.

In some embodiments, the PEGylation lipid compound is selected from at least one of PEG-modified phosphatidylethanolamine, PEG-modified phosphatidic acid, PEG-modified ceramide, PEG-modified dialkylamine, PEG-modified diacylglycerol, PEG-modified dialkylglycerol, and the above PEG-modified lipids modified with cell-targeting ligands.

In some embodiments, the delivery carrier further comprises an active ingredient, and the active ingredient is selected from at least one of DNA, RNA, a protein or a pharmaceutically active molecule.

In some embodiments, the delivery carrier is a lipid nanoparticle.

In some embodiments, the lipid nanoparticle further comprises an active ingredient, and the active ingredient is selected from at least one of DNA, RNA, a protein or a pharmaceutically active molecule.

In some embodiments, the RNA is selected from at least one of mRNA, siRNA, aiRNA, miRNA, dsRNA, aRNA, lncRNA, an antisense nucleotide (ASO) or an oligonucleotide.

In some embodiments, the protein is selected from at least one of an antibody, an enzyme, a recombinant protein, a polypeptide or a short peptide.

The present disclosure further provides a method for preparing lipid nanoparticles, the method comprising step (1) of mixing and dissolving the compound of the present disclosure, a PEGylation lipid, a structural lipid and a phospholipid in an absolute ethanol solution.

Optionally, the method further comprises step (2) of mixing the solution of the step (1) with an active ingredient to form lipid nanoparticles.

Optionally, the compound of the present disclosure, the PEGylation lipid, the structural lipid and the phospholipid are dissolved and mixed in ethanol, and then mixed with an active ingredient to form lipid nanoparticles.

In one embodiment, the present disclosure further provides the use of the compound of the present disclosure in the preparation of lipid nanoparticles.

In some embodiments, the compound of the present disclosure is selected from the following compounds, or salts or isomers thereof:

The present disclosure has the following beneficial effects:

When a numerical range is listed, each value and sub-range within the range are intended to be included. For example, “Calkyl” includes C, C, C, C, C, C, C, C, C, C, C, C, C, C, C, C, C, C, C, Cand Calkyl.

The term “alkyl” refers to a straight or branched saturated hydrocarbon group comprising one or more carbon atoms (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more carbon atoms). For example, “Calkyl” refers to an optionally substituted straight or branched saturated hydrocarbon group comprising 1-4 carbon atoms. “Calkyl” refers to an optionally substituted straight or branched saturated hydrocarbon group comprising 5-10 carbon atoms. Unless otherwise specified, the alkyl described herein refers to unsubstituted or substituted alkyl.